交联透明质酸植入材料在眼科应用的生物安全性评价  

Biological security evaluation of cross-linked hyaluronic acid for ophthalmic implant material

作　　者：郝念[1] 张惠成[1] 胡勇平[1] 刘晓玲[2] 董映[1] 

机构地区：[1]杭州市第一人民医院眼科,310006 [2]杭州师范大学医学实验中心,310016

出　　处：《中华实验眼科杂志》2013年第2期136-140,共5页Chinese Journal Of Experimental Ophthalmology

基　　金：浙江省科技计划项目（2008e23058）

摘　　要：【摘要】背景透明质酸是人体细胞外基质的一种黏多糖成分，具有良好的生物相容性。化学交联合成的交联透明质酸材料提高了材料的物理特性，具有临床应用前景。目的评价交联透明质酸材料的组织相容性及生物安全性，探讨其作为眼科植入材料的可行性。方法按照《医疗器械生物学评价》（GB／T16886．5—2003）的标准自行制备交联透明质酸材料。将18只8周龄雄性新西兰白兔采用随机配对法随机分为实验组和对照组，实验组兔眼角膜基质层间植入交联透明质酸材料3个月，对照组仅制作角膜基质层间囊袋，但不植入交联透明质酸材料，术后1周、2周，1、2、3个月裂隙灯显微镜观察交联透明质酸材料的在体角膜内生物学反应，并于术后2周、1个月、3个月行术眼角膜组织病理学检查。将小鼠胚胎成纤维细胞（L929细胞）分别置于消毒的交联透明质酸薄膜6孔细胞培养板（交联透明质酸组）、医用眼科硅胶材料6孔培养板（硅胶组）和未加任何材料6孔板内（阴性对照组），细胞培养48h后倒置显微镜及扫描电子显微镜观察L929细胞形态。L929细胞与交联透明质酸材料浸提液共同培养，48h后MTT法检测培养细胞的相对增生率（RGR）。结果兔在体实验的观察期内，交联透明质酸材料与兔角膜基质愈合良好，术后兔角膜无炎症反应和新生血管；组织病理学检查见角膜基质纤维排列整齐，未发现淋巴细胞等炎性细胞浸润。体外细胞接触试验后24h，交联透明质酸组和阴性对照组培养的细胞生长良好，细胞大小均一，排列正常，而硅胶组培养细胞贴壁生长欠佳，并有悬浮的死亡细胞。L929细胞与交联透明质酸材料浸提液共同培养48h后细胞的RGR为87．50％，属于细胞毒性1级。结论交联透明质酸材料在兔角膜组织中具有良好的组织相容性和生物安全性，是一种理�Background Hyaluronic acid is a mucopolysaccharide existing in extracellular matrix and having good biocompatibility. Using chemical crosslinking method can improve the physical properties of the material, so cross-linked hyaluronic has potential clinical application value. Objective The present study was to evaluate the histocompatibility and biological security of cross-linked hyaluronic acid as ophthalmic implant material. Methods Cross-linked hyaluronic acid implant material was prepared according to the criteria of Biological Evaluation of Medical Devices （ GB/T16886.5-2003 ）. Eighteen 8-week-old male New Zealand white rabbits were randomized into the experimental group and the control group. Cross-linked hyaluronie acid material with 5.0 mm diameter was implanted into corneal stroma interlaminationally in the experimental group, and only corneal stromal interlaminational pocket was made without any implanting material in the control group. Biological response of cornea was assessed in vivo from 1 week through 3 months after operation by slit lamp microscope. The corneas were obtained 2 weeks, 1 month and 3 months respectively for histopathological examination. Mouse embryonic fibroblasts were cultured in cross-linked hyaluronic acid film plate, medical silicone material culture plate and regular culture plate respectively for 24 hours, and the cell growth state and morphology were observed under the inverted microscope and scan electron microscope. MTT assay was used to test the relative growth rate of the cultured cells 48 hours after cultured using extracted liquid of hyaluronie acid implant material. Results Cross-linked hyaluronic acid implant material showed a well healing to the corneas of rabbits during the observation duration,without obvious inflammatory response and neovascularization. The arrangement of stromal fibers was uniform in order, and no infiltration of inflammatory cells was seen under the light microscope, The cells grew well after cultured with cross-linked hyaluronic acid film an

关 键 词：透明质酸 组织相容性 生物安全性 细胞毒性 

分 类 号：R318.08[医药卫生—生物医学工程]